Por favor, use este identificador para citar o enlazar a este item:
http://hdl.handle.net/10261/304734
COMPARTIR / EXPORTAR:
SHARE CORE BASE | |
Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL | DATACITE | |
Título: | A directed genome evolution method to enhance hydrogen production in Rhodobacter capsulatus |
Autor: | Barahona, Emma; Isidro, Elisa San; Sierra-Heras, Laura; Álvarez-Melcón, Inés; Jiménez-Vicente, Emilio; Buesa, José María; Imperial, Juan CSIC ORCID ; Rubio, Luis M. | Palabras clave: | Biological hydrogen production Flow cytometry HupA Hydrogenase Mutagenesis Nitrogenase |
Fecha de publicación: | 24-ago-2022 | Editor: | Frontiers Media | Citación: | Frontiers in Microbiology 13: e991123 (2022) | Resumen: | Nitrogenase-dependent H2 production by photosynthetic bacteria, such as Rhodobacter capsulatus, has been extensively investigated. An important limitation to increase H2 production using genetic manipulation is the scarcity of high-throughput screening methods to detect possible overproducing mutants. Previously, we engineered R. capsulatus strains that emitted fluorescence in response to H2 and used them to identify mutations in the nitrogenase Fe protein leading to H2 overproduction. Here, we used ultraviolet light to induce random mutations in the genome of the engineered H2-sensing strain, and fluorescent-activated cell sorting to detect and isolate the H2-overproducing cells from libraries containing 5 × 105 mutants. Three rounds of mutagenesis and strain selection gradually increased H2 production up to 3-fold. The whole genomes of five H2 overproducing strains were sequenced and compared to that of the parental sensor strain to determine the basis for H2 overproduction. No mutations were present in well-characterized functions related to nitrogen fixation, except for the transcriptional activator nifA2. However, several mutations mapped to energy-generating systems and to carbon metabolism-related functions, which could feed reducing power or ATP to nitrogenase. Time-course experiments of nitrogenase depression in batch cultures exposed mismatches between nitrogenase protein levels and their H2 and ethylene production activities that suggested energy limitation. Consistently, cultivating in a chemostat produced up to 19-fold more H2 than the corresponding batch cultures, revealing the potential of selected H2 overproducing strains. | Descripción: | 12 Pág. | Versión del editor: | https://doi.org/10.3389/fmicb.2022.991123 | URI: | http://hdl.handle.net/10261/304734 | DOI: | 10.3389/fmicb.2022.991123 | ISSN: | 1664-302X |
Aparece en las colecciones: | (INIA) Artículos |
Ficheros en este ítem:
Fichero | Descripción | Tamaño | Formato | |
---|---|---|---|---|
A_ directed_ genome_ evolution.pdf | artìculo | 2,19 MB | Adobe PDF | Visualizar/Abrir |
CORE Recommender
SCOPUSTM
Citations
3
checked on 09-may-2024
WEB OF SCIENCETM
Citations
3
checked on 25-feb-2024
Page view(s)
20
checked on 15-may-2024
Download(s)
61
checked on 15-may-2024
Google ScholarTM
Check
Altmetric
Altmetric
Este item está licenciado bajo una Licencia Creative Commons